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Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report date:
2015

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method

Test material

Constituent 1
Chemical structure
Reference substance name:
Citric acid, titanium salt
EC Number:
923-927-0
Molecular formula:
C30H32O35.Ti2
IUPAC Name:
Citric acid, titanium salt
Constituent 2
Chemical structure
Reference substance name:
Water
EC Number:
231-791-2
EC Name:
Water
Cas Number:
7732-18-5
Molecular formula:
H2O
IUPAC Name:
Dihydrogen oxide
Specific details on test material used for the study:
(1) Product name : CTL Ti638 UP
(2) CAS No. : Not available
(3) Lot No. : 110200/51
(4) Facility code No. : 2015/00032
(5) Received date : 26 May, 2015
(6) Appearance : Orange, particulate free liquid
(7) Purity : 54.7 wt.% (Ti content 4.97 wt.%)
(8) Storage condition : Room temperature
(9) Supplier : Catalytic Technologies Limited

Test animals

Species:
rat
Strain:
Sprague-Dawley
Remarks:
NTacSAM:SD
Sex:
female
Details on test animals or test system and environmental conditions:
Date of acquisition
- 1st acquisition : 25 June 2015
- 2nd acquisition : 10 July 2015
Number of animals received : Each 7 female rats per acquisition
Age of animals received : About 7 weeks
Body weights on arrival
- 1st acquisition : 174.89 - 189.04 g
- 2nd acquisition : 169.87 - 178.27
Quarantine and acclimation: Animals were acclimated for 5 (first step), 12 (second step), 6 (third step) and 13 (forth step) days. Only animals with the best appearance were selected for the test after observation during the acclimation period. Animals were accepted based on the certification provided by the supplier
Age at the initiation of the administration : About 8 - 9 weeks
Body weights at the administration
- G1 (1st step 300 mg/kg) : 183.50 - 185.36 g
- G2 (2nd step 300 mg/kg) : 180.02 - 194.07 g
- G3 (3rd step 2,000 mg/kg) : 181.95 - 184.09 g
- G4 (4th step 2,000 mg/kg) : 194.33 - 199.87 g
Number of animals administered: Three female rats were used in each step. The test substance was administered to total twelve female rats.
Grouping: Animals were weighed one day before the test substance administration and grouped to ensure a distribution of graded body weight. Animals whose body weights fall in an interval within ±20% of the mean weight are only used.

Environmental and Housing Condition
(1) Animal care room : Room 1 in the SPF animal facility area.
(2) Temperature and humidity : 23.6±0.9 °C and 58.3±3.6%RH
(3) Ventilation frequency : 10-15 air changes/hours
(4) Lighting cycle : 12 hours duration (lighting on at 8 a.m. and off at 8 p.m.)
(5) Lighting intensity : 281 Lux
(6) Noise : 56.2 dB
(7) Concentration of ammonia : less than 5 ppm
(8) Housing condition
All animals were housed in wire mesh cages (250W × 350L × 180H mm) during quarantine, acclimation, administration and observation period. During the experiment, not more than 3 animals were housed in a cage and cages were replaced at grouping.

Feeds and water
Radiation sterilized, solid laboratory animal feed (Teklad Certified Irradiated Global 18 % Protein Rodent Diet, Harlan Co. Ltd., USA) was provided ad libitum. DooYeol Biotech Co., Ltd. (91, Baumoe-ro, Seocho-gu, Seoul, Korea) supplied the feed. Tap water purified by reverse osmosis filtering system was provided ad libitum using water polycarbonate bottles.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Doses:
300 mg/kg and 2000 mg/kg
No. of animals per sex per dose:
3 females per dose
Details on study design:
Clinical signs and mortalities
General clinical signs or mortalities of all treated animals were carefully observed continuously during the first half-hour and the one hour from the
administration time. After that, all animals were observed regularly at one hour intervals until four hours on the administration day. From the next
day, animals were observed once every day up to day 14 after the administration. The administration day was regarded as day 0.

Body weight measurement
Individual animals were weighed at the date of acquisition, at grouping, dosing day prior to administration (on the administration day), on day 1, 7
and 14 (before necropsy) after administration.

Necropsy and gross findings examination
On 14 days after the administration, all survival animals were anesthetized with CO2 gas and terminated by exsanguination from the abdominal aorta
and caudal vena cava. Complete post-mortem examinations were performed on all vital organs.
Statistics:
Body weight changes of all animals in each step were analysed through tables and figures that were applied to the mean value and the standard
deviations. Additional statistical assessment was not performed in data analysis. The LD50 value was classified according to the study procedure diagram of OECD Guideline for the Testing of Chemicals No. 423 ‘Acute Toxic Class Method’ (Adopted 17th Dec., 2001).

Results and discussion

Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortalities were observed during the observation period at either dose level.
Clinical signs:
At 300 mg/kg no abnormal clinical signs were observed during the observation period.
At 2000 mg/kg 1/6 animal had inanimation and diarrhea on administration day. After that, emaciation, crawling position and paralytic gait were observed. In additon, black discoloration and hypoesthesia of distal parts of right foot and tail were revealed and those parts eventually were
lost. Hypoesthesia and paralytic gait were maintained until necropsy. Except that, there were no abnormal clinical signs in other animals.
Body weight:
At 300 mg/kg There were normal body weight gains in all animals.
At 2000 mg/kg temporary decrease was detected in 1/6 animal. Except that, there were normal body weight gains.
Gross pathology:
At 300 mg/kg no abnormal gross findings were observed.
At 2000 mg/kg loss of digits and distal part of tail were observed in 1/6 animal. Except that, no abnormal gross findings were observed.

Any other information on results incl. tables

Table 1. Mortalities and clinical signs of rats

SUMMARY OF MORTALITIES AND CLINICAL SIGNS

STUDY : GT15-00104

SEX : FEMALE

GROUP(/

G1(300)

G2(300)

G3(2,000)

G4(2,000)

MORTALITIES

N

0 / 3

0 / 3

0 / 3

0 / 3

%

0

0

0

0

CLINICAL SIGNS

No abnormalities detected

3 / 3

3 / 3

3 / 3

3 / 3

Diarrhea

0 / 3

0 / 3

1 / 3

0 / 3

Soiled perineal region

0 / 3

0 / 3

1 / 3

0 / 3

Inanimation

0 / 3

0 / 3

1 / 3

0 / 3

Emaciation

0 / 3

0 / 3

1 / 3

0 / 3

Black skin, distal part of foot, right

0 / 3

0 / 3

1 / 3

0 / 3

Black skin, distal part of tail

0 / 3

0 / 3

1 / 3

0 / 3

Crawling position

0 / 3

0 / 3

1 / 3

0 / 3

Paralytic gait

0 / 3

0 / 3

1 / 3

0 / 3

Hypoesthesia

0 / 3

0 / 3

1 / 3

0 / 3

Loss of tail, distal part

0 / 3

0 / 3

1 / 3

0 / 3

Loss of digits, right

0 / 3

0 / 3

1 / 3

0 / 3

Number of dead animals / Number of animals examined

Table 2. Body weight changes of rats

SUMMARY OF BODY WEIGHT CHANGES(g)

STUDY : GT15-00104

SEX : FEMALE

Day

GROUP(/)

G1(300)

G2(300)

G1(2,000)

G2(2,000)

 

0

184.45 ± 0.93 (3)

187.66 ± 7.11 (3)

182.84 ± 1.12 (3)

197.42 ± 2.82 (3)

1

208.72 ± 0.39 (3)

201.54 ± 8.19 (3)

191.78 ± 17.08 (3)

215.79 ± 15.55 (3)

7)

219.60 ± 4.01 (3

219.38 ± 8.96 (3)

202.75 ± 18.86 (3)

228.83 ± 10.08 (3)

14

241.00 ± 6.42 (3)

228.12 ± 12.18 (3)

215.33 ± 17.94 (3)

248.84 ± 10.86 (3)

Gaina

56.55 ± 5.59 (3)

40.46 ± 5.26 (3)

32.49 ± 17.56 (3)

51.42 ± 9.83 (3)

Mean±S.D (Number of animals)

a : Body weight gains between day 0 and 14

Applicant's summary and conclusion

Interpretation of results:
Category 5 based on GHS criteria
Conclusions:
The acute oral lethal dose 50 (LD50) of the test substance CTL Ti638 UP is considered greater than 2,000 ㎎/㎏ b.w. and it belongs to the GHS
category 5 under these study conditions.
Executive summary:

This study has been performed to evaluate the lethal dose 50 (LD50) and toxicity of the test substance CTL Ti638 UP when it was administered in single oral dose to Sprague-Dawley (SD) female rats. The test substance was administrated by gavage at the dose level of 300 ㎎/㎏ in two steps (the first and second step) and at the dose level of 2,000 ㎎/㎏ in two step (the third and forth step). During the study period, dead animals, clinical signs, body weight changes and gross findings at necropsy were examined.

1) In the first and second steps (300 ㎎/㎏), no mortalities and abnormal clinical signs were observed during the observation period. There were normal body weight gains in all animals. At the end of the study, necropsy was conducted to all animals and no abnormal gross findings were observed.

2) In the third and forth steps (2,000 ㎎/㎏), 1/6 animal had inanimation and diarrhea on administration day. After that, emaciation, crawling position and paralytic gait were observed. In additon, black discoloration and hypoesthesia

of distal parts of right foot and tail were revealed and those parts eventually were lost. Hypoesthesia and paralytic gait were maintained until necropsy. Except that, there were no abnormal clinical signs in other animals.

In body weights, temporary decrease was detected in 1/6 animal. Except that, there were normal body weight gains.

At the results of necropsy, loss of digits and distal part of tail were observed in 1/6 animal. Except that, no abnormal gross findings were observed.

Therefore, the acute oral lethal dose 50 (LD50) of the test substance CTL Ti638 UP is considered greater than 2,000 ㎎/㎏ b.w. and it belongs to the GHS category 5 under these study conditions.